Washed platelets were adjusted to 2 × 107/mL in Tyrode’s buffer (+ 0.35% BSA, + 2 mM CaCl2) and seeded on human fibrinogen coated coverslips (100 µg/mL) for 10 minutes before addition of agonists. Cells were fixed with 3.7% formaldehyde after 30 minutes, permeablized with 0.1% Triton X-100 and stained with phalloidin-594. Resting platelets were fixed in suspension and plated on poly-L-Lysine coated coverslips. Resting platelets were also stained with anti-β-tubulin (clone E7, Developmental Studies Hybridoma Bank, University of Iowa). Samples were analyzed on a Nikon Ti-U inverted microscope (Nikon Instruments Inc., Melville, NY) equipped with a Retiga EXL monochrome camera (QImaging, Surrey, Canada). Images were acquired using Slidebook 5.0 (Intelligent Imaging Innovations, Denver, CO) and a 100x oil objective. Cell surface area was quantified using Image J image analysis software.
Anti β tubulin clone e7
Manufactured by Developmental Studies Hybridoma Bank
Anti-β-tubulin (clone E7) is a monoclonal antibody that specifically recognizes the β-tubulin protein. It is a laboratory reagent used for the detection and study of β-tubulin in various applications.
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Lab products found in correlation
2 protocols using anti β tubulin clone e7
1
Visualizing Platelet Cytoskeleton Dynamics
2
Western Blot Analysis of Integrin Expression
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Pellets of fibroblasts were resuspended in Laemli Loading Buffer in reducing conditions, passed through a 23G needle 3 times and boiled for 5 min, fractionated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotted anti-β1 integrin (anti-CD29 clone 18 from BD Biosciences, dilution 1:1000), anti-GFP (Rockland 600-101-215, dilution 1:1000), or anti-β-tubulin (clone E7 from Developmental Studies Hybridoma Bank, dilution 1:1000) antibodies. After incubation with IRDye680 or IRDye800-conjugated secondary antibodies, the signal was scanned on an Odyssey CLx Imaging system, quantified and processed using Image Studio Lite (LI-COR Biosciences). Uncropped imaged are shown in Supplementary Figs. 10 and 11 .
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